Dear all,
I have some trouble for reverse transcription of high GC% content RNA (
800 bp, GC%=76% ).
Although I try to RT @ 65 degrees using ThermoSript RT system from BRL,
no PCR bands at all! However, with other high GC% template , I can
get the right size PCR product by adding DMSO into my reaction buffer .
So, the problem with my RT-PCR lies in RT step. Checking from gel, My
RNA is good before and after DNase I treatment.
Probably RT step contribute to my failure?
Does anyone has some tricks in RT such hard RNA ? Any suggestions will
be greatly appreciated.
Yang
UNC-CH, USA